Genus Lonomia is known for causing systemic poisoning from the inoculation of its poison through the skin, with consequent hemorrhagic manifestations with variable intensity, bringing the risk of death in some cases (Lorini, L. M., Passo Fundo, Brazil: EDIUPF, 1999, pages 25-35). The species Lonomia obliqua (Lemaire, C., Ann. Soc. Entomol. Fr. 8: 767-861, 1972) has caused epidemic accidents in areas of Southern Brazil (Ministério da Saúde, Fundação Nacional de Saúde, Acidentes por Lepidópteros in: Manual de diagnóstico e tratamento de acidentes por animais peçonhentos, Brasilia, 1998, page 131).
Patients who suffered accidents bear, among other symptoms, mainly after a period of 1 to 48 hours, blood dyscrasia (change in the proportion of blood elements), followed or not by hemorrhagical occurrences, which may cause death (Kelen, E. M. A. et al., J. Toxicol-Toxin Rev., 1995; 14: 283-308; Brazil, 1998).
Zannin established coagulation and fibrinolysis standards in plasma of 105 patients, and confirmed that poisoning affects coagulation and fibrinolysis. Their results showed intense coagulopathy consumption, which may be related to poison components in the bristles of caterpillars Lonomia obliqua, which have powerful procoagulant action, causing secondary activation of fibrinolysis (Zannin M. et al., Thromb. Haemost., 89: 355-364, 2003).
The extract of bristles of L. obliqua is effective for the experimental prevention of vein thrombosis in mice (Prezoto, B. C. et al., Braz. J. Med. Biol. Res. 2002; 35 (6): 703-12).
The poison of the caterpillar L. obliqua has some components which interfere in the coagulation system. The presence of prothrombin and Factor X activators in the extract of bristles of L. obliqua has been detected (Donato, J. L. et al., Thromb. Haemost. 1998; 79: 539-42; Kelen et al., 1995).
The authors of the invention have previously isolated and characterized a 69 kDa prothrombin activator protease called Lopap (Lonomia obliqua prothrombin activator protease), which has serinoprotease characteristics and procoagulant activity, exhausting blood of fibrinogen (Brazilian patent document PI 0200269). Lopap is structurally different from other prothrombin activators: the N-terminal portion bears 45.6% identity with the N-terminal portion of insecticianine of hemolymph of Manduca sexta; and Fragments I, II, III and IV show identity of 36.4%, 37.5%, 42.9% and 55.5%, respectively, with the corresponding internal fragments of insecticianine.
When intraperitoneally injected in mice in high concentrations (>100 μg/kg), Lopap develops thrombi in small veins and arteries, and the migration of polymorphonuclei to lungs and kidneys (Reis, C. V. et al., Lancet 1999, 353: 1942; Reis, C. V. et al., Thromb. Res. 2001, 102: 437-43; Reis, C. V. et al., Thromb. Res., 2001, 102: 427-436).
Lopap also acts on endothelial cells (HUVECs), as an expression inducer for adhesion molecules such as ICAM-1 and E-selectin, but not VCAM. The non-expression of VCAM suggests that the action of Lopap is not comparable to TNF-α or thrombin on endothelial cells.
The thrombin produced by Lopap is functional and inhibited by Antithrombin III (AT), being able to add platelets, coagulate plasma and fibrinogen, suggesting that this protease is similar to α-thrombin (Chudzinski-Tavassi, A. M. et al., Haemostasis 2001; 31: 257-265).
The recombinant form of Lopap is known. The Brazilian patent document PI 0403882 discloses a process to obtain recombinant Lopap (rLopap) in its monomeric form, its amino acid sequence, and its use as a defibrinogenating agent. The sequence of the recombinant protein, on average, presents 35% identity with lipocalin family proteins. Lipocalins are a family of proteins that store and transport hydrophobic and/or chemically sensitive organic compounds.